Name: generation of retinal organoids from healthy and retinal disease-specific human-induced pluripotent stem cells
Uploaded: 2022-12-09T13:40:00
Description: Watch this Scientific Journal Video about Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells at JoVE.com

作者感谢遗传学家Chitra Kannabiran博士的科学和技术支持;Subhadra Jalali博士，视网膜顾问;米林德·奈克博士，眼部整形外科医生;以及海得拉巴LV Prasad眼科研究所的眼部肿瘤学家Swathi Kaliki博士，致力于生成正常和患者特异性iPSC系。作者感谢科学与工程研究委员会，科学和技术部（IM）（SB / SO/HS / 177 / 2013），生物技术系（IM）（BT / PR32404 / MED / 30 / 2136 / 2019）的研发资助，以及来自ICMR（S.M.，D.P.），UGC（T.A.）和CSIR（V.K.P.）印度政府的高级研究奖学金。

所有涉及hiPSCs的实验均无菌进行，遵守标准实验室规范，伦理和生物安全指南，并得到机构伦理委员会（IEC），干细胞研究机构委员会（IC-SCR）和机构生物安全委员会（IBSC）等监管机构的批准。
1. iPSC培养和视网膜分化培养基及试剂的制备
<ol><li>iPSC 培养和维持培养基<ol><li>在无饲养层培养条件下，在基质胶包被（基底膜基质包被;参见材料表）培养?...

<ol>
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hiPSCs是体外研究器官和组织发育的有力工具。通过将健康与疾病特异性hiPSC与视网膜谱系区分开来概括疾病表型，有助于获得对不同形式遗传性视网膜营养不良的病理生理学的新见解。已经描述并采用了几种方案，用于将PSCs体外分化为视网膜细胞类型。其中大多数涉及使用含有重组生长因子、补充剂、小分子和试剂的复杂混合物的培养基，例如：N1、N2 和 B27 补充剂;BMP 和 TGFβ 信号阻...

视网膜是存在于脊椎动物眼睛后部的感光组织，它通过一种称为光转导途径的生化现象将光信号转化为神经冲动。视网膜感光细胞中产生的初始神经冲动被转导到其他视网膜中间神经元和视网膜神经节细胞（RGC）并到达大脑的视觉皮层，这有助于图像感知和视觉反应。
根据世界卫生组织（世卫组织）的数据，估计有150万儿童失明，其中100万在亚洲。遗传性视网膜营养不良症（IRD）是一种主要的致盲性疾病，全球每4，000人中就有1人受到影响1，2，3，而在发展中国家，与年龄相关性黄斑变性（AMD）相关的失明患病率在0.6%-1.1%之间4。IRD是由300多种不同基因的遗传缺陷引起的，这些基因涉及视网膜发育和功能5。这种遗传变化导致视网膜正常功能的破坏和视网膜细胞（即感光细胞和视网膜色素上皮（RPE））的逐渐退化，从而导致严重的视力丧失和失明。在涉及角膜、晶状体等的其他致盲疾病方面取得了巨大进展。然而，视网膜营养不良和视神经萎缩迄今没有任何经过验证的治疗方法。由于成人视网膜没有干细胞6，胚胎干细胞（ESC）和患者来源的诱导多能干细胞（iPSC）等替代来源可以提供无限供应所需的细胞类型，并为开发体外疾病建模研究和开发再生疗法所需的复杂组织类器官7提供了巨大的希望，8，9，10.
几年的视网膜研究使人们对协调早期视网膜发育的分子事件有了更好的理解。大多数从 PSC 生成视网膜细胞和 3D 类器官的方案旨在通过在生长因子和小分子的复杂混合物中培养细胞来逐步调节已知的生物过程，从而在体外概括这些发育事件。由此产生的视网膜类器官由主要的视网膜细胞组成：视网膜神经节细胞（RGC），中间神经元，光感受器和视网膜色素上皮（RPE）11，12，13，14，15，16，17，18，19.尽管成功尝试使用视网膜类器官对IRD进行建模，但在分化过程中对生长因子和小分子的复杂混合物的需求以及视网膜类器官生成效率相对较低，这对大多数方案构成了重大挑战。它们主要包括胚体的形成，然后在体外发育的不同阶段使用复杂的培养条件逐步分化为视网膜谱系20，21，22。
在这里，报道了一种从健康对照和视网膜疾病特异性hiPSCs开发复杂3D神经视网膜类器官的简化且稳健的方法。此处描述的方案利用近乎融合的hiPSC培养物的直接分化，而无需形成拟胚体。此外，培养基的复杂性也得到了简化，使其成为一种经济高效且可重复的技术，新研究人员可以轻松采用。它涉及一个混合培养系统，该系统由贴壁单层培养物组成，在视网膜分化的前 4 周内，直到出现独特的自组织眼野原始簇 （EFP）。此外，手动采摘每个EFP内的圆形神经视网膜岛并在悬浮培养物中生长1-2周，以制备由PAX6 +和CHX10 +增殖神经视网膜前体组成的多层3D视网膜杯或类器官。视网膜类器官在100μM含牛磺酸的培养基中进一步培养4周，导致RCVRN+和CRX+感光器前体的出现以及具有基本内段样延伸的成熟细胞。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#181;m Syringe filters</td>
			<td>TPP</td>
			<td>99722&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>15 mL centrifuge tube</td>
			<td>TPP</td>
			<td>91015</td>
			<td></td>
		</tr>
		<tr>
			<td>50 mL centrifuge tube</td>
			<td>TPP</td>
			<td>91050</td>
			<td></td>
		</tr>
		<tr>
			<td>6 well plates</td>
			<td>TPP</td>
			<td>92006</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Chx10 Antibody; Mouse monoclonal</td>
			<td>Santa Cruz</td>
			<td>SC365519</td>
			<td>1:50 dilution</td>
		</tr>
		<tr>
			<td>Anti-CRX antibody; Rabbit monoclonal</td>
			<td>Abcam</td>
			<td>ab140603</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Anti-MiTF antibody, Mouse monoclonal</td>
			<td>Abcam</td>
			<td>ab3201</td>
			<td>1:250 dilution</td>
		</tr>
		<tr>
			<td>Anti-Recoverin Antibody; Rabbit polyclonal&#160;&#160;&#160;&#160;&#160;</td>
			<td>Millipore</td>
			<td>AB5585</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>B-27 Supplement (50x), serum free</td>
			<td>Thermo Fisher</td>
			<td>17504044</td>
			<td></td>
		</tr>
		<tr>
			<td>Basic Fibroblast growth factor (bFGF)</td>
			<td>Sigma Aldrich</td>
			<td>F0291</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge 5810R</td>
			<td>Eppendorf</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Coplin Jar (50 mL)</td>
			<td>Tarson</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Corning Matrigel hESC-Qualified Matrix</td>
			<td>Corning</td>
			<td>354277</td>
			<td></td>
		</tr>
		<tr>
			<td>CryoTubes</td>
			<td>Thermo Fisher</td>
			<td>V7884</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEM/F-12, GlutaMAX supplement (basal medium)</td>
			<td>Thermo Fisher</td>
			<td>10565-018</td>
			<td></td>
		</tr>
		<tr>
			<td>DreamTaq DNA polymerase</td>
			<td>Thermo Fisher</td>
			<td>EP0709</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbeco&#8217;s Phosphate Buffered Saline</td>
			<td>Thermo Fisher</td>
			<td>14190144</td>
			<td></td>
		</tr>
		<tr>
			<td>Essential 8 medium kit</td>
			<td>Thermo Fisher</td>
			<td>A1517001</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethylene diamine tetraaceticacid disodium salt dihydrate (EDTA)</td>
			<td>Sigma Aldrich</td>
			<td>E5134</td>
			<td></td>
		</tr>
		<tr>
			<td>Falcon Not TC-treated Treated Petri Dish, 60 mm&#160;</td>
			<td>Corning</td>
			<td>351007</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal&#160;Bovine&#160;Serum, qualified, United States&#160;</td>
			<td>Gibco</td>
			<td>26140079</td>
			<td></td>
		</tr>
		<tr>
			<td>GelDocXR+ with Image lab software</td>
			<td>BIO-RAD</td>
			<td></td>
			<td>Agarose Gel documentation system&#160;</td>
		</tr>
		<tr>
			<td>GlutaMAX Supplement</td>
			<td>Thermo Fisher</td>
			<td>35050061</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-Mouse IgG (H+L), Alexa Fluor 488</td>
			<td>Invitrogen</td>
			<td>A11001</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Goat anti-Mouse IgG (H+L), Alexa Fluor 546</td>
			<td>Invitrogen</td>
			<td>A11030</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Goat anti-Rabbit IgG (H+L), Alexa Fluo 546</td>
			<td>Invitrogen</td>
			<td>A11035</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Goat anti-Rabbit- IgG (H+L), Alexa Fluor 488</td>
			<td>Invitrogen</td>
			<td>A11008</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>HistoCore MULTICUT</td>
			<td>Leica</td>
			<td></td>
			<td>For sectioning</td>
		</tr>
		<tr>
			<td>KnockOut Serum Replacement</td>
			<td>Thermo Fisher</td>
			<td>10828028</td>
			<td></td>
		</tr>
		<tr>
			<td>L-Acsorbic acid</td>
			<td>Sigma Aldrich</td>
			<td>A92902</td>
			<td></td>
		</tr>
		<tr>
			<td>MEM Non-Essential Amino Acids Solution (100x)</td>
			<td>Thermo Fisher</td>
			<td>11140-050</td>
			<td></td>
		</tr>
		<tr>
			<td>N2 supplement (100x)</td>
			<td>Thermo Fisher</td>
			<td>17502048</td>
			<td></td>
		</tr>
		<tr>
			<td>NanoDrop 2000</td>
			<td>Thermo Fisher</td>
			<td></td>
			<td>To quantify RNA</td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Qualigens</td>
			<td>23995</td>
			<td></td>
		</tr>
		<tr>
			<td>Pasteur Pipets, 9 inch, Non-Sterile, Unplugged</td>
			<td>Corning</td>
			<td>7095D-9</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin&#160;</td>
			<td>Thermo Fisher</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant Anti-Otx2 antibody , Rabbit monoclonal</td>
			<td>Abcam</td>
			<td>ab183951</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Recombinant Anti-PAX6 antibody; Rabbit Monoclonal</td>
			<td>Abcam</td>
			<td>ab195045</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Recombinant Anti-RPE65 antibody, Rabbit Monoclonal</td>
			<td>Abcam</td>
			<td>ab231782</td>
			<td>1:300 dilution</td>
		</tr>
		<tr>
			<td>Recombinant Human Noggin Protein</td>
			<td>R&#38;D Systems</td>
			<td>6057-NG</td>
			<td></td>
		</tr>
		<tr>
			<td>SeaKem LE Agarose</td>
			<td>Lonza</td>
			<td>50004</td>
			<td></td>
		</tr>
		<tr>
			<td>Serological pipettes 10 mL</td>
			<td>TPP</td>
			<td>94010</td>
			<td></td>
		</tr>
		<tr>
			<td>Serological pipettes 5 mL</td>
			<td>TPP</td>
			<td>94005</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>S7653</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Citrate Tribasic dihydrate</td>
			<td>Sigma Aldrich</td>
			<td>S4641</td>
			<td></td>
		</tr>
		<tr>
			<td>Starfrost (silane coated) microscopic slides</td>
			<td>Knittel</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>SuperScript III First-Strand Synthesis System</td>
			<td>Thermo Fisher</td>
			<td>18080051</td>
			<td></td>
		</tr>
		<tr>
			<td>SuperScript III First-Strand Synthesis System for RT-PCR</td>
			<td>Invitrogen</td>
			<td>18080051</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma Aldrich</td>
			<td>T8787</td>
			<td></td>
		</tr>
		<tr>
			<td>TRIzol&#160;Reagent</td>
			<td>Invitrogen</td>
			<td>15596026</td>
			<td></td>
		</tr>
		<tr>
			<td>UltraPure 0.5 M EDTA, pH 8.0</td>
			<td>Thermo Fisher</td>
			<td>15575020</td>
			<td></td>
		</tr>
		<tr>
			<td>VECTASHIELD Antifade Mounting Medium with DAPI&#160;</td>
			<td>Vector laboratories</td>
			<td>H-1200&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Vitronectin</td>
			<td>Thermo Fisher</td>
			<td>A27940</td>
			<td></td>
		</tr>
		<tr>
			<td>Y-27632 dihydrochloride (Rho-kinase inhibitor)</td>
			<td>Sigma Aldrich</td>
			<td>Y0503</td>
			<td></td>
		</tr>
		<tr>
			<td>Zeiss LSM 880</td>
			<td>Zeiss</td>
			<td></td>
			<td>Confocal microscope</td>
		</tr>
	</tbody>
</table>

generation of retinal organoids from healthy and retinal disease-specific human-induced pluripotent stem cells

多能干细胞可以产生复杂的组织类器官，可用于体外疾病建模研究和开发再生疗法。该协议描述了一种更简单，稳健和逐步的方法，该方法在视网膜分化的前4周内在由贴壁单层培养物组成的杂交培养系统中生成视网膜类器官，直到出现独特的，自组织的眼场原始簇（EFP）。此外，每个EFP内的甜甜圈形，圆形和半透明神经视网膜岛在视网膜分化培养基中使用非粘附培养皿在悬浮液下手动挑选和培养1-2周以产生多层3D光学杯（OC-1M）。这些未成熟的视网膜类器官含有PAX6+和ChX10+增殖的多能视网膜前体。前体细胞在类器官内线性自组装，表现为明显的径向条纹。悬浮培养后4周，视网膜祖细胞经历有丝分裂后停滞和谱系分化，形成成熟的视网膜类器官（OC-2M）。光感受器谱系承诺的前体在视网膜类器官的最外层发育。这些CRX+和RCVRN+感光细胞形态成熟，显示出内段样延伸。该方法可用于使用人胚胎干细胞（hESCs）和诱导多能干细胞（iPSCs）生成视网膜类器官。所有步骤和程序都得到清晰的解释和演示，以确保可复制性并在基础科学和转化研究中得到更广泛的应用。

hiPSCs分化为眼谱系是通过在不同时间点依次在含有补充剂和生长因子的不同培养基混合物中培养细胞来实现的，如图1所示。hiPSC培养物维持在Essential 8培养基（多能干细胞维持培养基）中。一旦它们达到70%-80%的汇合度（图2A），在第0天（参考步骤3.2）用含有1 ng / mL bFGF，1 ng / mL Noggin和1% N 2添加剂的分化诱导培养基（DIM）替换培养基。与神经诱导的N2补?...

Watch this Scientific Journal Video about Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells at JoVE.com

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

该协议描述了一种将hiPSCs分化为眼场簇并使用涉及贴壁和悬浮培养系统的简化培养条件生成神经视网膜类器官的有效方法。其他眼细胞类型，如RPE和角膜上皮，也可以在视网膜培养物中从成熟的眼视野中分离出来。

从健康和视网膜疾病特异性人诱导的多能干细胞中生成视网膜类器官

Pluripotent stem cells can generate complex tissue organoids that are useful for in vitro disease modeling studies and for developing regenerative ...

该协议描述了一种将人类IPC分化为复杂的三维神经视网膜类器官以进行研究和再生应用的简单有效的方法。该技术涉及贴壁和悬浮培养，允许选择性挑选和富集视网膜类器官和RP细胞。它可以为开发基于细胞的视网膜退行性疾病疗法提供可行且定期的细胞源供应。这种干细胞来源的视网膜类器官和RPE细胞可作为体外模型用于研究眼睛发育和遗传性视网膜疾病。这种方法可以很容易地被已经熟悉处理人类IPSC培养物的研究实验室复制。视觉演示极大地支持了眼野的识别和基于其空间定位和形态特征的神经视网膜杯的分离。首先，在六孔板中吸出70%至80%融合的人IPSC培养物的用过的培养基。将PBS添加到孔中，旋转它们并吸出洗涤缓冲液。然后向每个孔中加入一毫升细胞解离溶液或CDS，并将板在37摄氏度下孵育五到七分钟，直到细胞四舍五入。吸出CDS并将细胞悬液收集到15毫升离心管中。在室温下以 1， 000 RPM 旋转管子四分钟。弃去上清液并将细胞沉淀重悬于1.2毫升Essential 8培养基中。将200微升细胞悬液分配到含有1.5毫升IPSC培养基和10微摩尔Y27632的基质包被的六孔板的每个孔中。12至24小时后，除去用过的培养基并加入不含Y27632的预热完全提升培养基以维持培养物。每24小时更换一次培养基，直到培养物达到70%至80%汇合度。在第零天，吸出人IPSC维持培养基，并向六孔板中加入含有每毫升BFGF一毫微克和每毫升Noggin一毫微克的分化诱导培养基。将细胞在37摄氏度下在5%二氧化碳培养箱中孵育。在第一天，吸出用过的培养基并加入含有每毫升 1 纳克 BFGF 和 10 纳克/毫升 Noggin 的分化诱导培养基。在37摄氏度的5%二氧化碳培养箱中再次孵育细胞。在第 2 天和第 3 天，取出用过的培养基并加入含有每毫升 10 ngg Noggin 的分化诱导培养基。将每孔培养基加入两毫升到六孔板后，将细胞在37摄氏度下在5%二氧化碳培养箱中孵育，每24小时更换一次培养基。在第四天，去除用过的培养基并添加视网膜分化培养基或RDM。按照相同的步骤将每孔培养基加入两毫升到六孔板中并孵育培养物。每天更换培养基。在第14天和第18天左右，在显微镜下以10倍放大倍率观察培养物，以发现由早期眼野祖细胞组成的神经莲座状结构域的出现。在第 21 天和第 28 天左右，在显微镜下以 4X 和 10X 放大倍率观察培养物的出现，以观察自组织的不同 EFP 的出现，其中心岛由圆形 3D 神经视网膜结构组成，周围环绕着连续的神经上皮和眼表上皮生长。取无菌巴斯德移液管，一只手握住底座，另一只手握住毛细管尖端。通过旋转运动对毛细管尖端中间附近的区域进行火焰消毒和加热，直到玻璃变得柔韧。然后远离火焰，迅速向外拉毛细管尖端，形成具有封闭管腔的细毛细管尖端。将细尖端水平握在火焰前面，然后以向外运动快速通过火焰，以形成光滑的钩子或 L 形毛细管尖端。在体视显微镜下，使用毛细管钩的光滑外曲率区作为细勺，从各个EFP中手动挑选形状良好的神经视网膜杯。在第 25 天和第 30 天，在低附着 60 毫米培养皿中加入 4 毫升预热的视网膜分化培养基，以培养和维持视网膜杯以生成 3D 视网膜类器官。这应该在收获神经视网膜杯之前完成。将一毫米微量移液器设置为吸出 100 微升，并使用具有宽孔开口的一毫升微量吸头吸出并将浮动视网膜杯转移到之前准备的新鲜低贴壁培养皿中。将视网膜杯保持在RDM中作为非贴壁悬浮培养物，并在37%二氧化碳培养箱中以37摄氏度孵育它们。在第30天和第45天，遵循部分进料方法，去除一半体积的废培养基，并用等体积的新鲜培养基代替。在第45天和第60天，在含有100微摩尔牛磺酸的RDM中再培养视网膜类器官两周，以支持神经视网膜祖细胞的更好存活和谱系分化以及成熟视网膜细胞类型的发育。视网膜类器官在不同的成熟阶段表征，用于使用RT-PCR和免疫组织化学表达几种视网膜祖细胞标志物。RT-PCR结果证实了一个月大的视网膜类器官和两个月大的视网膜类器官中神经视网膜标志物的诱导和表达。此处显示了使用针对神经视网膜祖细胞标志物 Chx10、PAX6 和 Otx2 的抗体的未成熟视网膜类器官的免疫标记切片的共聚焦图像，以及使用针对感光器前体标志物恢复蛋白和 CRX 的抗体的成熟视网膜类器官的共聚焦图像。具有分化感光细胞的视网膜类器官的标记外层被放大并显示在这些图像中。基本的内段状延伸部分用白色箭头标记。以神经视网膜罩为中心且迁移的 RPE 生长物的 EFP 簇沿前缘显示色素沉着。此处显示了神经视网膜岛周围多个EFP的分化良好的色素上皮生长。EFP 的较高放大倍率显示 RPE 祖细胞的迁移区和神经视网膜杯周围的眼表上皮。此处显示了发展出含有色素和非色素细胞的未成熟RPE细胞单层的扩展贴壁培养物。完全成熟和色素沉着的RPE细胞的单层培养物在第60天显示出鹅卵石形态。表达PAX6，MITF和RPE65的RPE细胞如图所示。这些图像代表具有异常聚集体的视网膜祖细胞的EFP，具有扭曲的层压和缺乏条纹。EFP与微型神经视网膜杯，但缺乏RPE和眼表上皮的周围区域如图所示。代表性图像显示了悬浮培养物中形成的不规则神经视网膜聚集体。使用接近融合的IPC培养物开始分化过程非常重要，以便在三到四周内实现眼野的有效诱导。由正常和患者特异性IPC产生的视网膜类器官和RPE细胞可用作视网膜疾病模型和测试新型治疗药物。

Research

JoVE Journal

Developmental Biology

Efficient Derivation of Retinal Pigment Epithelium Cells from Stem Cells

视网膜色素上皮细胞的干细胞高效的推导

No cure has been discovered for age-related macular degeneration  (AMD), the leading cause of vision loss in people over the age of 55. AMD is complex ...

科研

发育生物学

Deriving Retinal Pigment Epithelium (RPE) from Induced Pluripotent Stem (iPS) Cells by Different Sizes of Embryoid Bodies

Deriving Retinal Pigment Epithelium RPE from Induced Pluripotent Stem iPS Cells by Different Sizes of Embryoid Bodies

从诱导多能干得出视网膜色素上皮（RPE）（IPS），由胚体大小不同的细胞

Pluripotent stem cells possess the ability to proliferate indefinitely and to differentiate into almost any cell type. Additionally, the development of ...

Generation of Integration-free Human Induced Pluripotent Stem Cells Using Hair-derived Keratinocytes

集成的无人类诱导多能干细胞的生成方法头发衍生角质形成细胞

Recent advances in reprogramming allow us to turn somatic cells into human induced pluripotent stem cells (hiPSCs). Disease modeling using ...

Three-dimensional Quantification of Dendritic Spines from Pyramidal Neurons Derived from Human Induced Pluripotent Stem Cells

树突棘从锥体神经元的三维定量从人类诱导多能干细胞衍生

Dendritic spines are small protrusions that correspond to the post-synaptic compartments of excitatory synapses in the central nervous system. They are ...

Defined and Scalable Generation of Hepatocyte-like Cells from Human Pluripotent Stem Cells

界定和由人多能干细胞的肝细胞样细胞的可扩展代

Human pluripotent stem cells (hPSCs) possess great value for biomedical research. hPSCs can be scaled and differentiated to all cell types found in the ...

Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells

分化软骨细胞与外周血源性人诱导多能干细胞

In this study, we used peripheral blood cells (PBCs) as seed cells to produce chondrocytes via induced pluripotent stem cells (iPSCs) in an ...

Rapid, Directed Differentiation of Retinal Pigment Epithelial Cells from Human Embryonic or Induced Pluripotent Stem Cells

人胚胎或诱导多能干细胞对视网膜色素上皮细胞的快速定向分化

We describe a robust method to direct the differentiation of pluripotent stem cells into retinal pigment epithelial cells (RPE). The purpose of providing ...

Generation of Standardized and Reproducible Forebrain-type Cerebral Organoids from Human Induced Pluripotent Stem Cells

人诱导多能干细胞的前脑型脑 Organoids 标准化和可再生性的生成

The human cortex is highly expanded and exhibits a complex structure with specific functional areas, providing higher brain function, such as cognition. ...

Generation of 3D Skin Organoid from Cord Blood-derived Induced Pluripotent Stem Cells

从脐带血源性多能干细胞中提取三维皮肤有机细胞

The skin is the body&#8217;s largest organ and has many functions. The skin acts as a physical barrier and protector of the body and regulates bodily ...

In Vitro Generation of Somite Derivatives from Human Induced Pluripotent Stem Cells

人诱导多能干细胞体外生成体细胞衍生物的研究

In response to signals such as WNTs, bone morphogenetic proteins (BMPs), and sonic hedgehog (SHH) secreted from surrounding tissues, somites (SMs) give ...